Resinas acrílicas utilizadas em placas estabilizadoras: resistência a fraturas e ensaio microbiológico / Acrylic resins used in stabilizer plates: resistance to fractures and microbiological testing

O aprimoramento da tecnologia CAD/CAM permitiu a confecção de placas estabilizadoras digitais, no entanto, há lacunas acerca do comportamento desses dispositivos frente a variáveis mecânicas e biológicas ainda desconhecidas. Dessa forma, o objetivo desse trabalho foi verificar se as resinas utilizadas na tecnologia digital das placas fresadas e impressas são capazes de influenciar na resistência à fratura e na adesão de micro-organismos em detrimento das resinas empregadas na confecção das placas termopolimerizáveis convencionais e se as propriedades testadas se relacionam com a variação da espessura desses dispositivos. Para tanto, foi realizado um estudo in vitro que contou com as fases mecânica e microbiológica, sendo cada uma composta pelos grupos Convencional (GC) (n=30), Fresado (GF) (n=30) e Impresso (GI) (n=30), de acordo com o método de confecção, possuindo cada um três subgrupos (n=10) que variaram a espessura dos espécimes de 1 a 3 mm. Na fase mecânica, as amostras, com formato de barras de 65 mm, foram testadas quanto à resistência à fratura na máquina de ensaios universal, com célula de carga de 500 kgf e velocidade de 1 mm/min; na microbiológica, a forma foi de discos com 15 mm de diâmetro, submetidos à adesão de micro-organismos à superfície com a exposição a Streptococcus mutans e mantidos por 24h a 35 °C em estufa para sucessiva contagem de UFC, com controle positivo. Os testes foram feitos em triplicata e os dados foram armazenados no SPSS 22.0 e a análise estatística contou com a ANOVA e o pósteste de Tukey. Nos resultados da fase mecânica, o ANOVA identificou diferenças estatisticamente significativas entre os grupos (p≤0,005), mostrando que GC e GF apresentaram melhores resultados. O pós-teste de Tukey considerou diferença estatisticamente significativa entre GF e GI (p=0,031), elencando GF como superior. Para a fase microbiológica, não foram verificadas diferenças estatísticas entre os grupos (p>0,005) em nenhuma das análises. Observou-se que para a resistência a fraturas, as resinas do modo fresado descreveram resultados superiores e/ou próximos aos da técnica convencional; e que as resinas da técnica digital e convencional, quanto à adesão superficial de micro-organismos, não diferiram entre si (AU).

CAD/CAM technology allowed the stabilizing splints manufacture, however there are gaps about these devices' behavior in face of mechanical and biological variables that are still unknown. Thus, the objective of this project was to verify if the resins used in digital technology of milled and printed stabilizer splints can influence fracture resistance and microorganism adhesion, in detriment of the resin applied in manufacture of conventional thermopolymerizable splints and if the tested properties are related to thickness variation of these devices. The method included mechanical and microbiological steps, each one composed by Conventional (CG) (n=30), Milled (MG) (n=30) and Printed (PG) (n=30) groups, according to the method of preparation, each one having three subgroups (n=10) that varied the specimen thickness from 1 to 3 mm. In the mechanical step, the samples, in the form of 65 mm bars, were tested for fracture resistance in a universal testing machine, with load cell of 500 kgf and speed of 1 mm/min; in the microbiological step, the shape was in form of 15 mm diameter discs, subjected to microorganism adhesion to the surface with exposure to Streptococcus mutans and kept for 24 hours at 35 °C in oven for successive CFU counting, with positive control. The tests were performed in triplicate and data were stored in SPSS 22.0 and statistical analysis used ANOVA and Tukey's post hoc. In the results of mechanical step ANOVA had identified statistically significant differences between the groups (p≤0,005), showing CG and MG had presented better results. Tukey´s post hoc had considered statistically significant difference between MG and PG (p=0,031), listing MG as superior. For the microbiological step, they did not show statistical differences between the groups (p>0.005) in any analysis. It was observed milled resins described superior results and/or close to the conventional technique one, for fracture resistance; and resins of digital and conventional technique, regarding the microorganism adhesion to the surface, did not differ from each other (AU).

Manipulation of Focal Adhesion Signaling by Pathogenic Microbes.

Focal adhesions (FAs) serve as dynamic signaling hubs within the cell. They connect intracellular actin to the extracellular matrix (ECM) and respond to environmental cues. In doing so, these structures facilitate important processes such as cell-ECM adhesion and migration. Pathogenic microbes often modify the host cell actin cytoskeleton in their pursuit of an ideal replicative niche or during invasion to facilitate uptake. As actin-interfacing structures, FA dynamics are also intimately tied to actin cytoskeletal organization. Indeed, exploitation of FAs is another avenue by which pathogenic microbes ensure their uptake, survival and dissemination. This is often achieved through the secretion of effector proteins which target specific protein components within the FA. Molecular mimicry of the leucine-aspartic acid (LD) motif or vinculin-binding domains (VBDs) commonly found within FA proteins is a common microbial strategy. Other effectors may induce post-translational modifications to FA proteins through the regulation of phosphorylation sites or proteolytic cleavage. In this review, we present an overview of the regulatory mechanisms governing host cell FAs, and provide examples of how pathogenic microbes have evolved to co-opt them to their own advantage. Recent technological advances pose exciting opportunities for delving deeper into the mechanistic details by which pathogenic microbes modify FAs.

A post-invasion role for Chlamydia type III effector TarP in modulating the dynamics and organization of host cell focal adhesions.

The human pathogen Chlamydia trachomatis targets epithelial cells lining the genital mucosa. We observed that infection of various cell types, including fibroblasts and epithelial cells resulted in the formation of unusually stable and mature focal adhesions that resisted disassembly induced by the myosin II inhibitor, blebbistatin. Superresolution microscopy revealed in infected cells the vertical displacement of paxillin and focal adhesion kinase from the signaling layer of focal adhesions, whereas vinculin remained in its normal position within the force transduction layer. The candidate type III effector TarP, which localized to focal adhesions during infection and when expressed ectopically, was sufficient to mimic both the reorganization and blebbistatin-resistant phenotypes. These effects of TarP, including its localization to focal adhesions, required a post-invasion interaction with the host protein vinculin through a specific domain at the C terminus of TarP. This interaction is repurposed from an actin-recruiting and -remodeling complex to one that mediates nanoarchitectural and dynamic changes of focal adhesions. The consequence of Chlamydia-stabilized focal adhesions was restricted cell motility and enhanced attachment to the extracellular matrix. Thus, via a novel mechanism, Chlamydia inserts TarP within focal adhesions to alter their organization and stability.

Cocoon-like fibroadhesive tuberculous peritonitis in a peritoneal dialysis patient.

Tuberculous peritonitis is a devastating complication of peritoneal dialysis (PD). Presentations of tuberculous peritonitis range from the common wet ascitic form to the rare fibroadhesive form, which is clinically indistinguishable from encapsulating peritoneal sclerosis. We describe a 76-year-old man on continuous ambulatory PD for three months developing wet ascitic form of tuberculous peritonitis. Three weeks after institution of antituberculous therapy and removal of PD catheter, his abdominal pain recurred and an encapsulating peritoneal sclerosis-like intestinal obstruction was noted. A rare fibroadhesive form of tuberculous peritonitis associated with the paradoxical response to antituberculous therapy was considered by excluding noncompliance, drug resistance and adverse effects, and other concomitant infections. After surgical enterolysis and continuation of antituberculous treatment, he recovered uneventfully. Our case might be the first report regarding paradoxical deterioration to antituberculous treatment in dialysis patients.

When Shigella tells the cell to hang on.

OspE, a Shigella type III effector binds to integrin-like kinase and enhances cell adhesion to better disseminate and colonize the intestinal epithelium. Because of the existence of OspE orthologues in other enteropathogens such as enteropathogenic Escherichia coli or Salmonella sp., maintenance of cell adhesion appears as a widespread strategy for bacteria that interact with the intestinal epithelium.

Identification of residues in the N-terminal domain of the Yersinia tyrosine phosphatase that are critical for substrate recognition.

YopH is a 468-amino acid protein-tyrosine phosphatase that is produced by pathogenic Yersinia species. YopH is translocated into host mammalian cells via a type III protein secretion system. Translocation of YopH into human epithelial cells results in dephosphorylation of p130(Cas) and paxillin, disruption of focal adhesions, and inhibition of integrin-mediated bacterial phagocytosis. Previous studies have shown that the N-terminal 129 amino acids of YopH comprise a bifunctional domain. This domain binds to the SycH chaperone in Yersinia to orchestrate translocation and to tyrosine-phosphorylated target proteins in host cells to mediate substrate recognition. We used random mutagenesis in combination with the yeast two-hybrid system to identify residues in the YopH N-terminal domain that are involved in substrate-binding activity. Four single codon changes (Q11R, V31G, A33D, and N34D) were identified that interfered with binding of the YopH N-terminal domain to tyrosine-phosphorylated p130(Cas) but not to SycH. These mutations did not impair YopH translocation into HeLa cells infected with Yersinia pseudotuberculosis. Introduction of the V31G substitution into catalytically inactive (substrate-trapping) forms of YopH interfered with the ability of these proteins to bind to p130(Cas) and to localize to focal adhesions in HeLa cells. In addition, the V31G substitution reduced the ability of catalytically active YopH to dephosphorylate target proteins in HeLa cells. These data indicate that the substrate- and SycH-binding activities of the YopH N-terminal domain can be separated and that the former activity is important for recognition and dephosphorylation of substrates by YopH in vivo.